Polish Journal of Veterinary Sciences Vol. 22, No. 1 (2019), 17–24 DOI 10.24425/pjvs.2018.125596 Original article Efficacy of β-hydroxy-β-methylbutyric acid (HMB) for growing rate and its influence for health indicators in blood test of young early-weaning goats 1K. Cebulska, 1P. Sobiech, 2S. Milewski, 2K. Ząbek 1 Department of Internal Disease, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Oczapowskiego 14, 10-957 Olsztyn 2 Department of Sheep and Goat Breeding, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-917 Olsztyn Abstract The study was conducted on 26 male, 30 days-old goats, separated from their mothers, divided into two equal groups: I - control and II - experimental, consisting of 13 animals each. All animals were fed with milk replacer, experimental group received additionally 50 g/kg body weight, additive of HMB, for 60 days. The following features were analyzed: body weight, daily increases of body weight, as well as hematological and biochemical blood features. Differences in body weight were found, between experimental and control group, after 60 days of experiment 0.57 kg (p≤0.01). The daily weight gain of experimental animals was higher in comparison with control group. Significant differences were also noted in results of hematological and biochemical blood parameters. Experimental animals showed a higher level of red blood cells as well as number of lymphocytes in comparison with the control group, (p≤0.01).Significant changes were also observed in the level of triglycerides, inorganic phosphorus and protein between both groups. The acid-base balance parameters – – and ionogram, showed a higher pH level (p≤0.05) HCO3 act., HCO3 std., BE, ctCO2, O2sat, K+, Cl– (p≤0.01), while the anion gap (AG) and Na+ were significantly lower in control group (p≤0.01). Key words: small ruminant, kids, feed additives, muscle mass, hematology, biochemistry Correspondence to: K. Ząbek, e-mail:[email protected] 18 K. Cebulska et al. Introduction effect of HMB should also be noted. Stimulating effect of this compound was observed in cellular and humoral Production of goats milk is associated with the ne- defense of broilers chicken (Peterson et al. 1999) and cessity of early separation of young animals from their fish (Siwicki et al. 2000, 2001, 2003). The authors of mothers and such situation may negatively affect their mentioned studies noticed on increase of phagocytic future growth (Szymanowska et al. 1997). Development capacity of macrophages, lymphocyte proliferation of effective breeding technology, demands creating bal- both by ConA (concanavalin A) and LPS (lipopolysac- anced feeding patterns and it is a significant research charide), and higher level of ASC (antibody secreting problem. The substance which can stimulate the animal cells). growth, in such circumstances is β-hydroxy-β-methylbu- HMB plays a positive role in animal organism. Con- tyric acid (HMB) (Tatara et al. 2007, Tatara 2008, sidering fact that the influence of HMB on growth and Tatara et al. 2012). health of early-weaned goats has not been studied so Endogenous HMB is one of the leucine metabolites far, the aim of this research was to investigate HMB as (Van Koevering and Nissen 1992). There is also a pos- a diet supplement, its effect on health status and growth sibility to merge exogenous HMB, produced by chemi- of early-weaned goats. cal or microbiological synthesis into metabolic path- ways (Van Koevering and Nissen 1992, Vukovich et al. 2001). In humans, HMB is used as a diet supplement, Materials and Methods which supports the muscular cells activity and strength – in case of intense workouts or in patients suffering The study was planned and conducted in accor- from cachexia, AIDS or terminal cancer (Fitschen et al. dance to Ethical Committee Approval, decision No. 2013). The inhibitory effect of HMB on apoptotic pro- 42/2014, 25.06.2014. The experiment was conducted on cesses, has also been demonstrated in human and 26 male, Alpine goats, from single births. chicken muscle cells (Kornasio et al. 2009). Muscle mass gain, improving the nitrogen balance and hemato- Experimental design logical parameters, was observed after using HMB in Goats weaned from mothers at the age of 30 ± 3 diet (Jowko et al. 2001, Portal et al. 2002, Nissen and days were divided into two groups, consisting 13 ani- Sharp 2003, Flakoll et al. 2004, Rathmacher et al. 2004, mals in each: I - control and II - experimental. Animals Kuhls et al. 2007). from both groups had a similar mean body weight at the HMB plays also an important role in protein me- beginning of the study (6,40 kg for control group and tabolism by inhibiting proteolysis, supporting protein 6,43 for experimental group). During the 60 days of and cholesterol synthesis (Bloch et al. 1954, Adamson research, all animals received the same diet: the milk and Greenberg 1955, Bachhawat et al. 1955, Gey et al. replacer WITAMILK 2 (Table 1), produced by Wipasz 1957, Ostaszewski et al. 2000, Smith et al. 2005). Olsztyn (Poland) at a dose of 1.5 l /for one animal/day, Scientific research demonstrated the stimulating with water and grass haylage ad libitum. Animals from effect of HMB in muscle development in turkeys experimental group additionally received, β-hydroxy- (Moore et al. 2005). One of the theories says, that these -β-methylbutyric acid (HMB), with a supplemental changes are associated with stimulation of intestinal mixture, at dose of 50 mg/kg of body weight per day. villi development (Tako et al. 2004), which leads to Throughout the experiment, the amount of consumed increased forage intake. Research conducted by Tatara and non-consumed forage was controlled. Chemical et al. (2007) indicate another mechanism that may composition was determined using standard methods be associated with neuroendocrine regulators, such as (AOAC 2005). The individual food intake, in both growth hormone (GH) and insulin-like growth factor 1 groups, was determined during the entire experiment (IGF-1), both in humans and animals. An increased on basis of the results obtained. level of these factors was observed in the offspring The animals were kept in accordance with the of sows receiving HMB during their pregnancy (Tatara guidelines included in the animal protection act, were et al. 2007, 2012). On the other hand, such changes under the constant veterinarian supervision and they were not observed in broilers (Qiao et al. 2013). Previ- didn’t show any symptoms that could suggest illness. ous studies indicated that HMB takes part in increasing the level of certain amino-acids in organism of young lambs (Papet et al. 1997), as well as the improvement of bone system of lambs (Krupski et al. 2011). In addition to the positive effect of HMB on increasing the muscle mass, the immunomodulatory Efficacy of β-hydroxy-β-methylbutyric acid (HMB) ... 19 Table 1. Components of WITAMILK 2 produced final body weight – initial body weight by Wipasz Olsztyn (Poland). RGR = x 100 (%) Ingredients WITAMILK 2 1/2 (initial body weight + final body weight) Whole protein % 20.00 Hematological and biochemical blood analyses Crude fat % 10.00 Crude fiber % 0.80 Blood for analyses was sampled from external jugu- NaCl % 1.40 lar vein three times: before the beginning of the exper- Na % 0.60 iment (day 0) and on the 30th and 60th day. Ca % 0.70 Hematological tests included: white blood cell count (WBC), leucogram determination (including: P org. % 0.60 percentage of basophils, eosinophils, granulocytes and K org. % 1.50 segmented granulocytes, lymphocytes and monocytes), Methionine % 0.40 red blood cell count (RBC), hematocrit value (HCT), Lactose % 30.00 hemoglobin level (HGB), mean corpuscular volume Vit A Ul 50 000.00 (MCV), mean corpuscular hemoglobin (MCH), mean Vit D3 Ul 8 000.00 corpuscular hemoglobin concentration (MCHC) and Vit E mg 100.00 platelet count (PLT). These parameters were deter- Vit K3 mg 4.00 mined in whole blood using a Siemens hematology an- Vit B1 mg 10.00 alyzer ADVIA 2120. Biochemical tests included: glucose, total proteins, Vit B2 mg 8.00 cholesterol, triglycerides (TG), inorganic phosphorus Nicotinic acid mg 40.00 (Pinorg), calcium (Ca), magnesium (Mg), ionogram – Pantothenic acid mg 30.00 Na+, K , Cl+, acid – base balance – pH, partial pres- Vit B6 mg 8.00 sure of carbon dioxide and oxigen (pCO and pO ), bi- Vit B12 mg 0.05 carbonate⁺ concentration (HCO –), base excess (BE), Biotin mg 0.10 oxygen saturation (O sat), CO concentration₂ (ctCO₂ ) 2 ₃ Choline chloride mg 500.00 and anion gap. This parameters were determined using ₂ ₂ Vit C mg 150.00 following methods: enzymatic method (glucose, choles- Folic acid mg 0.95 terol level) colorimetric method (total protein, trigly- cerides, Ca, P , Mg level ). Parameters were obtained Mg % 0.20 inorg with the Cormay ACCENT 200, using Cormay diagnos- Fe mg 120.00 tic kits. Mn mg 50.00 Parameters of acid-base balance and electrolytes Cu mg 15.00 in venous blood were determined using the RapidLab Zn mg 90.00 348 from Siemens. I mg 1.20 Hematological and biochemical blood parameters Co mg 2.00 were analyzed in laboratories of the Faculty of Veteri- Se mg 0.30 nary Medicine University of Warmia and Mazury, Olsztyn, Poland. Probiotic -/+ + Statistical analyzes Analyzed parameters The results of the study were statistically analyzed using the analysis of variance (ANOVA) for univariate The growth rate systems (body mass and daily increases) and two-factor systems (blood indicators). Significance of differences Analyses of lambs growth included determinations between groups was verified using the post-hoc Duncan of their body weight at the beginning of the experiment, test. The mean values presented in Tables 4-6 are given after 30 and 60 days, as well as daily body weight gains together with the standard error (± SE).